42P - Biological role of prognostic microRNAs (miRNAs) in squamous lung carcinoma (SCC) cells

Date 17 April 2015
Event ELCC 2015
Session Poster lunch
Topics Lung and other Thoracic Tumours
Translational Research
Presenter Martyna Filipska
Citation Annals of Oncology (2015) 26 (suppl_1): 10-14. 10.1093/annonc/mdv045
Authors M. Filipska1, M. Skrzypski2, G. Stasiłojć1, J. Bigda3
  • 1Department Of Medical Biotechnology, Medical University of Gdansk, 80-211 - Gdansk/PL
  • 2Department Of Oncology And Radiotherapy, Medical University of Gdansk, Gdansk/PL
  • 3Department Of Medical Biotechnology, Medical University of Gdansk, Gdansk/PL

Abstract

Aim/Background

To date, there are no reliable tools allowing for individual selection of high-risk non-small cell lung cancer (NSCLC) patients for adjuvant chemotherapy. We earlier demonstrated that overexpression of three miRNAs: miR-662, -192 and -192* may correlate with the risk of distant relapse in SCC patients undergoing pulmonary resection (Skrzypski et al. 2014, Br J Cancer). However, predictive value of these miRNAs for chemosensitivity of SCC is unknown. The aim of this study was to assess the biological role of three abovementioned miRNAs in SCC cells.

Methods

In a search for appropriate in vitro model we screened by reverse transcription quantitative PCR (RT-qPCR) 11 NSCLC cell lines for miRNA expression profiles and assessed their sensitivity for cisplatin and etoposide by MTT cytotoxic test. Candidate cell lines were further analyzed in migration, invasion and soft agar clonogenic assays. Cells naturally overexpressing miR-662, -192 and -192* were transfected with locked nucleic acid (LNATM) miRNA inhibitors. The functionality of these inhibitors was first confirmed by GFP reporter gene assay. The inhibitor-treated and WT cells were compared in cytotoxic tests and soft agar colony formation assay.

Results

Among analyzed NSCLC cell lines we found a SCC cell line with natural overexpression of miR-662, -192 and -192*. These cells were resistant to chemotherapeutics and exhibited an ability to form colonies in soft agar. The inhibition of miR-192* and miR-662 sensitized SCC cells for cisplatin (p = 0.032 and p = 0.024, respectively). Moreover, inhibition of miR-192 and miR-662 resulted in reduction of colony number in comparison to negative control (p < 0.001).

Conclusions

We developed and optimized a cellular model for in vitro study of biological role of three prognostic miRNAs in SCC. Initial cytotoxic tests showed influence of miR-192* and miR-662 on cisplatin chemosensitivity, we therefore plan to continue a search for mechanisms maintained by these miRNAs in SCC. Our results also suggest that genes targeted by miR-192 and/or miR-662 may be involved in maintaining the clonogenic potential of SCC. Further studies may elucidate predictive value of these genes.

Disclosure

All authors have declared no conflicts of interest.